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LSRII filters on a CSU-X1 confocal head

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Benoit Labarthe

LSRII filters on a CSU-X1 confocal head

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello, flowers and microscopists!

Here is a chimera-type question!

We have emission filters from an BD LSRII flow cytometer (disks of 25mm of diameter) and would like to use them in a CSU-X1 confocal head. Basically, the filters are fitting in the emission wheel of the CSU and we see fluorescence. My question is: do emission filters for flow cytometry and for confocal microscopy have the same properties?
I may be wrong, but both are the final emission filters in front of either the PMT or the camera and thus have a similar role of filtering the emissions. In my mind, both filters should be comparable. Do I omit any critical detail?

Thank you!

benoit

--
Benoît Labarthe, PharmD
Institut de Cardiologie de Montréal
Montreal Heart Institute
Centre de Recherche, Laboratoire du Dr.Théroux
5000 Bélanger Est
Montréal Québec Canada H1T 1C8
Tel (514)-376 3330 ext 3017
Fax (514)- 376 1076

Guy Cox

Re: LSRII filters on a CSU-X1 confocal head

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I don't know of any differences. I'm sure you'll have no problems.

It is usually a good idea to keep the filters the same way round

(some have an arrow to indicate the direction the light should

go). So the side facing the PMT in the flow cytometer should

be the side facing the camera in the microscope.

Guy

Optical Imaging Techniques in Cell Biology
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Electron Microscope Unit, Madsen Building F09,
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Subject: LSRII filters on a CSU-X1 confocal head

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Matt Burton-2

Re: LSRII filters on a CSU-X1 confocal head

In reply to this post by Benoit Labarthe

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Benoît,

the filters are the same, there should be no problem using them. Note the
15.8 mm dichroic mirrors used in a LSR II have an AOI of 11.25 degrees not
the standard 45.

Matt

At 01:51 PM 13/03/2008 Thursday, you wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello, flowers and
>microscopists!
>
>Here is a chimera-type question!
>
>We have emission filters from an BD LSRII flow cytometer (disks of 25mm of
>diameter) and would like to use them in a CSU-X1 confocal head. Basically,
>the filters are fitting in the emission wheel of the CSU and we see
>fluorescence. My question is: do emission filters for flow cytometry and
>for confocal microscopy have the same properties?
>I may be wrong, but both are the final emission filters in front of either
>the PMT or the camera and thus have a similar role of filtering the
>emissions. In my mind, both filters should be comparable. Do I omit any
>critical detail?
>
>Thank you!
>
>benoit
>
>--
>Benoît Labarthe, PharmD
>Institut de Cardiologie de Montréal
>Montreal Heart Institute
>Centre de Recherche, Laboratoire du Dr.Théroux
>5000 Bélanger Est
>Montréal Québec Canada H1T 1C8
>Tel (514)-376 3330 ext 3017
>Fax (514)- 376 1076

Michael Weber-4

Re: LSRII filters on a CSU-X1 confocal head

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

CSU-X1 is not using 45 degree dichomatic mirrors either, they also have
low-angle incidence ones. I am not sure about the precise angle, however,
these mirrors have to be produced and there are probably not so many
different low-angles out there (for Nikon, Zeiss, Yokogawa). But it might
make sense to ask Yokogawa for the correct angle.

Michael

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Benoît,
>
> the filters are the same, there should be no problem using them. Note the
> 15.8 mm dichroic mirrors used in a LSR II have an AOI of 11.25 degrees not
> the standard 45.
>
> Matt
>
> At 01:51 PM 13/03/2008 Thursday, you wrote:
>>Search the CONFOCAL archive at
>>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello, flowers
>> and
>>microscopists!
>>
>>Here is a chimera-type question!
>>
>>We have emission filters from an BD LSRII flow cytometer (disks of 25mm
>> of
>>diameter) and would like to use them in a CSU-X1 confocal head.
>> Basically,
>>the filters are fitting in the emission wheel of the CSU and we see
>>fluorescence. My question is: do emission filters for flow cytometry and
>>for confocal microscopy have the same properties?
>>I may be wrong, but both are the final emission filters in front of
>> either
>>the PMT or the camera and thus have a similar role of filtering the
>>emissions. In my mind, both filters should be comparable. Do I omit any
>>critical detail?
>>
>>Thank you!
>>
>>benoit
>>
>>--
>>Benoît Labarthe, PharmD
>>Institut de Cardiologie de Montréal
>>Montreal Heart Institute
>>Centre de Recherche, Laboratoire du Dr.Théroux
>>5000 Bélanger Est
>>Montréal Québec Canada H1T 1C8
>>Tel (514)-376 3330 ext 3017
>>Fax (514)- 376 1076

Mizuho Shimizu

Re: LSRII filters on a CSU-X1 confocal head

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear List,

Please let me clarify CSUX1 filter requirements.

As for emission filters, you can use any commercailly available emission filters of 25mm diameter as long as they have appropriate wavelength range and enough blocking capability.

However, the dichroic (dichromatic) mirrors for the CSU heads(all CSU10/22/X1) have reverse charcteristics from those used for standard microscope.
Please imagine the location of the dichroic mirror in the CSU scanners, which is placed between the microlens array and pinhole array at about 45 degree angle. The excitation laser beamlets focused by the the microlens will pass through the dicroic mirror and then go through the pinholes. The emission light goes back the light path to the pinholes then reflrcted by the dichroic mirror to reach at the detector(CCD camera). So, please only use specific dichroic mirrors for the CSU heads. Also, the thichness of the dichroic mirrors should be 0.5 mm +/- 0.02mm(0.01mm is more desirable) for optimal light efficiency.

Please always feel free to directly contact Yokogawa for any questions.

Best regards,

Mizuho Shimizu
***********************************************************
Mizuho Shimizu
CSU Business Development Group
BIO Center
Life Science Business Headquarters
Yokogawa Electric Corporation
2-3 Hokuyodai, Kanazawa-shi, Ishikawa
920-0177 JAPAN
Phone: +81-76-258-7028 Fax: +81-76-258-7029
http://www.yokogawa.co.jp/SCANNER/english/index_e.html

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差出人: Confocal Microscopy List [[hidden email]] は Michael Weber [[hidden email]] の代理
送信日時: 2008年3月14日 16:56
宛先: [hidden email]
件名: Re: LSRII filters on a CSU-X1 confocal head

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

CSU-X1 is not using 45 degree dichomatic mirrors either, they also have
low-angle incidence ones. I am not sure about the precise angle, however,
these mirrors have to be produced and there are probably not so many
different low-angles out there (for Nikon, Zeiss, Yokogawa). But it might
make sense to ask Yokogawa for the correct angle.

Michael